HOPE-Fixation of Lung Tissue Allows Retrospective Proteome and Phosphoproteome Studies

Shevchuk, Olga; Abidi, Nada; Klawonn, Frank; Wissing, Josef; Nimtz, Manfred; Kügler, Christian; Steinert, Michael; Goldmann, Torsten; Jänsch, Lothar

doi:10.1021/pr500096a

Cited by 5 publications

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References 48 publications

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“…HOPE® fixation technique (HEPES-glutamic acid buffer mediated Organic solvent Protection Effect) (DCS Diagnostics, Hamburg, Germany) was used as previously published [ 39 , 40 ]. There were no washing steps for the ex vivo infected tissues before fixation.…”

Section: Methodsmentioning

confidence: 99%

Mycobacteria infect different cell types in the human lung and cause species dependent cellular changes in infected cells

et al. 2016

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BackgroundMycobacterial infections remain a significant cause of morbidity and mortality worldwide. Due to limitations of the currently available model systems, there are still comparably large gaps in the knowledge about the pathogenesis of these chronic inflammatory diseases in particular with regard to the human host. Therefore, we aimed to characterize the initial phase of mycobacterial infections utilizing a human ex vivo lung tissue culture model designated STST (Short-Term Stimulation of Tissues).MethodsHuman lung tissues from 65 donors with a size of 0.5–1 cm3 were infected each with two strains of three different mycobacterial species (M. tuberculosis, M. avium, and M. abscessus), respectively. In order to preserve both morphology and nucleic acids, the HOPE® fixation technique was used. The infected tissues were analyzed using histo- and molecular-pathological methods. Immunohistochemistry was applied to identify the infected cell types.ResultsMorphologic comparisons between ex vivo incubated and non-incubated lung specimens revealed no noticeable differences. Viability of ex vivo stimulated tissues demonstrated by TUNEL-assay was acceptable. Serial sections verified sufficient diffusion of the infectious agents deep into the tissues. Infection was confirmed by Ziel Neelsen-staining and PCR to detect mycobacterial DNA. We observed the infection of different cell types, including macrophages, neutrophils, monocytes, and pneumocytes-II, which were critically dependent on the mycobacterial species used. Furthermore, different forms of nuclear alterations (karyopyknosis, karyorrhexis, karyolysis) resulting in cell death were detected in the infected cells, again with characteristic species-dependent differences.ConclusionWe show the application of a human ex vivo tissue culture model for mycobacterial infections. The immediate primary infection of a set of different cell types and the characteristic morphologic changes observed in these infected human tissues significantly adds to the current understanding of the initial phase of human pulmonary tuberculosis. Further studies are ongoing to elucidate the molecular mechanisms involved in the early onset of mycobacterial infections in the human lung.

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Section: Methodsmentioning

confidence: 99%

Mycobacteria infect different cell types in the human lung and cause species dependent cellular changes in infected cells

et al. 2016

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“…In addition to the two reviews that update the current knowledge , and a technical development for clinical tissue preservation, a wide variety of human diseases were investigated in this collection of proteome studies. These include diseases of central nervous system, − eye, , lung, − breast, − oral cavity, esophagus, , liver, − pancreas, colorectum, − ovarian, − cervix, muscle, , skin, , blood cells, circulatory system, , and kidney. , In addition, infection and immunity − (including Ebola virus infection with the current and largest outbreak in history that affects multiple countries, particularly those in West Africa, and triggers worldwide attention), and metabolic disorders − were also reported.…”

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Recent Advances of Proteomics Applied to Human Diseases

Thongboonkerd

LaBaer²,

Domont

2014

J. Proteome Res.

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G iven the intimate roles proteins play in health and disease, it is not surprising that during the past 20 years proteomics has been applied extensively to investigate human diseases to better understand disease mechanisms/pathways and to define new therapeutic targets as well as novel biomarkers for accurate diagnostics/prognostics. There is no doubt that advancements of disease proteomics will expand the frontiers of medicine to impact global health.This Special Issue of Journal of Proteome Research (JPR) features recent progress and advancements of proteomic studies of human diseases, focusing on pathogenesis, pathogenic mechanisms, diagnosis, prognosis, and treatment. It includes two reviews and 64 research articles (a total of 66 articles). The large number of articles published in a single Special Issue reflects the significance and broad interest of this area.It is especially encouraging to use the occasion of this Special Issue to reflect on how our field has evolved. Compared with disease proteomics articles previously published, this collection of studies shows obvious improvements in this field as follows. First, more advanced proteomic technologies, particularly sophisticated mass spectrometry for protein characterizations, profiling, quantitative analysis, and investigations of posttranslational modifications (PTMs), were used. Second, the study designs were more sophisticated and planned more carefully, for example, stringent validation to confirm screening data sets, sufficient sample size, and suitable control groups (i.e., inclusion of diseased controls in addition to the normal control). Third, functional significance of the identified proteins or targets was confirmed in most of these studies. Finally, feasibility for clinical applications was well demonstrated, for example, sensitivity/specificity of novel biomarkers for diagnostics/prognostics.In addition to the two reviews that update the current knowledge 1,2 and a technical development for clinical tissue preservation, 3 a wide variety of human diseases were investigated in this collection of proteome studies. These include diseases of central nervous system, 4−14 eye, 15,16

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Infection of Human Lung Tissue Explants (HLTEs) with Legionella pneumophila

Scheithauer

Steinert

2019

Methods in Molecular Biology

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HOPE-Fixation of Lung Tissue Allows Retrospective Proteome and Phosphoproteome Studies

Cited by 5 publications

References 48 publications

Mycobacteria infect different cell types in the human lung and cause species dependent cellular changes in infected cells

Mycobacteria infect different cell types in the human lung and cause species dependent cellular changes in infected cells

Recent Advances of Proteomics Applied to Human Diseases

Infection of Human Lung Tissue Explants (HLTEs) with Legionella pneumophila

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